Molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in China.
Molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in China.
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DOI：
10.1007/s11262-007-0196-7
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发表时间：
2008-04
期刊：
影响因子：
1.6
通讯作者：
Feng L
中科院分区：
文献类型：
--
作者：
Chen JF;Sun DB;Wang CB;Shi HY;Cui XC;Liu SW;Qiu HJ;Feng L
从2006年我国感染猪流行性腹泻病毒（PEDV）的仔猪粪便中分离到6株PEDV。采用RT-PCR方法扩增6个PEDV分离株的膜蛋白基因，克隆、测序，并与10个PEDV参考株进行比较。6个PEDV中国分离株的M蛋白基因全长692个核苷酸，含有一个681个核苷酸的开放阅读框，编码一个226 aa的多肽。在6个PEDV分离物的M蛋白基因起始端ATG上游5个核苷酸处发现了与Br 1/87相同的保守基因间基序（ATAAAC）。在CV 777、JMe 2、LZC和QH中也发现了六聚体基序。6个分离物的M蛋白具有3个主要的跨膜结构域（aa 20 -38、aa 43 -65、aa 75 -97）。CH/IMT/06的M蛋白有一个潜在的糖基化位点，而其他5个分离株的M蛋白有两个潜在的糖基化位点。糖基化序列Asn-Phe-Thr在6个PEDV分离物的M蛋白中高度保守。6个PEDV分离物的核苷酸序列同源性为98.8 ~ 100%，推导的氨基酸序列同源性为98.2 ~ 100%。6个PEDV分离株与10个参考株的M蛋白基因核苷酸和氨基酸同源性分别为97.2 ~ 99.4%和96.9 ~ 100%。根据M蛋白基因的系统发育关系，6个PEDV分离株组成一个独立的簇，其中包括一个中国株JS-2004-02，但不包括中国株LJB/03。这些结果表明，我国存在一种新的PEDV基因型。
Six porcine epidemic diarrhea viruses (PEDVs) were isolated from the fecal samples of piglets infected with PEDV in 2006 in China. The membrane (M) protein genes of six PEDV isolates were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR), then cloned, sequenced, and compared with each other as well as those ten PEDV reference strains. The M protein genes of six Chinese PEDV isolates consisted of 692 nucleotides containing a single open reading frame (ORF) of 681 nucleotides, which encoded a 226aa-long peptide. The conserved intergenic motif (ATAAAC), as previously recognized in Br1/87, was found in the 5 nucleotides upstream of the initiator ATG of M protein genes of six Chinese PEDV isolates. The hexamer motif was also found in CV777, JMe2, LZC, and QH. The M protein of six isolates had three main transmembrane domains (aa20–38, aa43–65, aa75–97). The M protein of one isolate, CH/IMT/06, had one potential glycosylation site, but those of the other five isolates had two. The glycosylation sequence Asn-Phe-Thr was highly conserved in the M proteins of six PEDV isolates. The six PEDV isolates showed nucleotide sequence homology between 98.8 and 100% and deduced amino acid sequence homology between 98.2 and 100% with each other. The nucleotide and amino acid identity of M protein genes between the six PEDV isolates and ten reference PEDV strains varied from 97.2 to 99.4% and 96.9 to 100%, respectively. On the basis of the phylogenetic relationship of M protein genes, six Chinese PEDV isolates composed of a separate cluster including one Chinese strain JS-2004-02, however, not including the Chinese strain LJB/03. These results demonstrated that there was a new genotype of PEDV prevailing in China.
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